| (19) |
 |
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(11) |
EP 0 731 176 A3 |
| (12) |
EUROPEAN PATENT APPLICATION |
| (88) |
Date of publication A3: |
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24.11.1999 Bulletin 1999/47 |
| (43) |
Date of publication A2: |
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11.09.1996 Bulletin 1996/37 |
| (22) |
Date of filing: 07.03.1996 |
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| (51) |
International Patent Classification (IPC)6: C12Q 1/68 |
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| (84) |
Designated Contracting States: |
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AT BE CH DE ES FR GB IT LI |
| (30) |
Priority: |
07.03.1995 JP 74616/95
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| (71) |
Applicant: NIPPON ZOKI PHARMACEUTICAL CO., LTD. |
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Chuo-ku,
Osaka (JP) |
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| (72) |
Inventor: |
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- Imai, Hideki,
c/o Inst. of Bio-Active Science
Yashiro-cho,
Katoh-gun,
Hyogo (JP)
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| (74) |
Representative: Kolb, Helga, Dr. Dipl.-Chem. et al |
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Hoffmann, Eitle & Partner,
Patent-und Rechtsanwälte,
Arabellastrasse 4
81925 München
81925 München (DE) |
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| |
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| (54) |
Method of gene duplication and amplification |
(57) An object of the present invention is to offer a method for the duplication and also
for the amplification of gene wherein the base sequence is ambiguous.
The constitution of the present invention is a method for duplication of gene, characterized
in that, DNA which is complementary to RNA or double-stranded DNA of gene wherein
the base sequence is ambiguous is dissociated into single-stranded DNA, then primer
is partially bonded to a nonspecific position of the single-stranded DNA, DNA moiety
having a complementarity to the single-stranded DNA is synthesized by DNA polymerase
using said bonded primer as a starting point of the DNA synthesis and the above-mentioned
operation is repeated twice or more whereby the nonspecific moiety of the gene is
duplicated and also is a method of amplification of said gene using a PCR method thereafter.
The merit of the present invention is that gene wherein the base sequence is ambiguous
can be duplicated and amplified and that it can be utilized for elucidation of unknown
pathogenic microorganisms, for diagnosis of said infectious disease and for development
of vaccine therefor.